This is a Phase I SBIR proposal to test the feasibility of using an ultra miniaturized protein NanoArray for profiling cytokine expression in mouse model systems. BioForce Nanosciences, Inc., has developed proprietary technology for the construction of NanoArrays/TM, an ultra miniaturized assay platform that allows a thousand or more molecular tests to be carried out in the same surface space occupied by a single standard microarray spot. Using this technology, we propose to build a 9 component antibody sandwich NanoArray and test its sensitivity, precision and linearity using mouse recombinant cytokines and cell culture model systems. The size reduction offered by the NanoArray platform, relative to the conventional microarray format, provides a mechanism for the creation of diagnostic tests that require extremely small sample volumes, approaching that of a single cell. This assay system is particularly well-suited for studies involving small animal model systems, where conventional analysis techniques may require lethal amounts of starting sample material. We predict that our system will provide robust protein profiling data from mouse tail-bleeds, needle biopsy aspirates, and finger pricks of human blood samples. Furthermore, the NanoArray platform provides the benefits of miniaturization but still allows readout using standard fluorescence methods permitting data collection from a single image. This attribute increases the availability of array-based bioanalyses for scientists with limited budgets and/or instrumentation resources, and opens the doorway for clinical testing not currently possible with existing methodology. We have chosen cytokine biomarkers for our Phase I study. Cytokines have been implicated in the pathogenesis of a wide variety of diseases of the kidney and digestive systems, including diabetes, progressive renal failure and inflammatory bowel disease. Using the cytokine benchmark will allow us to produce a NanoArray biomarker screen that can be compared with existing standard ELISA and microarray-based tests, and be immediately useful to researchers using transgenic mouse models of disease. Development of this assay will pave the way for Phase 11development of protein profiling chips tailored to other disease targets and/or signaling pathways, for use in mouse model systems, and for human testing involving extremely small sample volumes.